Home | Archive | AJA @ Nature | Online Submission | News & Events | Subscribe | APFA | Society | Links | Contact Us | 中文版 |
 |
Cytokines
in the BALB/c mouse testis in various conditions
E.
Veräjänkorva1, M. Martikainen1,
P. Pöllänen1,2
Department
of Anatomy1 and Department of Obstetrics and Gynecology2,
University of Turku, FIN-20520 Turku, Finland
Asian J Androl 2001 Mar; 3: 9-19
Keywords:
testosterone;
vasectomy; estrogens; cryptorchidism; abdominal testis; varicocele;
cyokines; IL-10; macrophages; clonal anergy
Abstract
Aim: To investigate whether testosterone, estrogens, vasectomy, experimental cryptorchidism, varicocele or aging would induce changes in the cytokine environment of the mouse testis. Methods: In adult male BALB/c mice, testosterone implants, estradiol benzoate, vasectomy, unilateral cryptorchidism, unilateral varicocele were administered/performed. The mice were followed up for different periods of time and were then sacrificed with testes incised for examination. The control mice received the vehicle or sham-operation. Results: IL-10 was present in Leydig cells of nearly every testis and IL-10+ macrophages in 39% of testes. IL-6 was found in the testes of intact adult mice, mice treated with testosterone for 70 days, cryptorchid testes and sham-operated testes. Conclusion: Results suggest that IL-10 might be involved in the generation of the immunologically privileged microenvironment in the testis.
1 Introduction
Testis
is an immunologically privileged site where nearly every pathological
condition of the male reproductive tract is associated with an increased
prevalence of sperm
antibodies[1]. The cytokines have two major roles in the testis:
1) to mediate pathophysiological
outcomes of immune-endocrine interactions during inflammatory disease,
2) to work as growth and differentiation factors that help to orchestrate
cellular interactions during normal physiological functions[2].
It is known that the cells responsible for the maintenance of the blood-testis
barrier in the testis, the Sertoli cells, produce an IL-1-like factor[3,4]
and IL-6[5-7], as well as TGF[8-10] and Fas Ligand[11].
In addition, it has been reported that IL-11[12] is expressed
in differentiating male germ cells. IFN- and -[13] and
TNF-[14] are produced by testicular cells. However, a comprehensive
study on the cytokine environment of the testis in physiological as well
as pathological conditions has not yet been made.
The
cells mainly responsible for protection of germ cells from autoimmune
reactions, the Sertoli cells and the principal cells of the epididymis,
are dependent on androgens[15,16], the concentration of which
is decreased in both the
testis and the epididymis during male hormonal contraception[17].
A decrease in androgen
level may result in the breakdown of blood-testis and blood-epididymis
barrier, the release of sperm autoantigens and the development of autoimmune
destruction of the male reproductive tract. Indeed, it has been reported
that the blood-testis barrier starts to break down in conditions of subnormal
testosterone production in the seasonal breeder mink[16] with
the formation of sperm antibodies[18]. In such a process, cytokines
should have a distinctive role. This is also true in case of disrupting
spermatogenesis in cryptorchidism[19] and
vasectomy[20,21]. In all these conditions, the microenvironment
of the testis might have been changed.
2 Materials and methods
2.1
Animals
BALB/c
male mice 2 (prepubertal), 4 (pubertal), 11 and 25 weeks of age were used as
donors of normal testicular tissue. Each group consisted of at least three
mice. The animals had free access to food and water and they were maintained
in a normal dark/light
cycle. Permissions for the experiments and to use organs from the
animals after sacrifice with CO2 (Permission 659/96) were granted
by the local animal authorities.
2.2
Antibodies
Monoclonal
antibodies against mouse cytokines were used as primary antibodies. The
antibodies were as follows: rat-anti-mouse IL-2 (IgG2a; Pharmingen,
clone S4B6), rat-anti-mouse IL-4 (IgG1; Pharmingen, clone 11B11),
rat-anti-mouse IL-6 (IgG1; Pharmingen, clone MP5-20F3), rat-anti-mouse
IL-10 (IgG2b; Pharmingen, clone JES5-16E3), rat-anti-mouse
IL-12 (IgG1; Pharmingen, clone C15.6, reactive to the p40
subunit), rat-anti-mouse TNF- (IgG1; Pharmingen, clone MP6-XT3),
rat-anti-mouse IFN- (IgG1; Pharmingen, clone R4-6A2) and
rat anti-mouse CD106 (VCAM-, c
2.3
Administration of testosterone
To
simulate the male hormonal contraception, a subcutaneous silastic (medical
grade tubing, ID 3.35 mm, OD 4.65 mm, Aromando Medizintechnik, Dsseldorf,
Germany; closed with Medical adhesive silicone type A, Dow Corning Medical
Products, Midland, Michigan, USA) testosterone implant (one cm in length)
or an empty (control) implant was introduced through a small incision
under the back skin of 12 normal, over 11-weeks old male mice under pentobarbital
anaesthesia (60 mg/kg, Mebunat, Orion, Finland). The wound was closed
with 5-0 Dermalon (American Cyanamid company, Wayne, NJ, USA) sutures.
The mice were followed for 17, 35, 52 and 70 days,
after which they were sacrificed with CO2. The testes were
collected and frozen in liquid nitrogen for later use.
2.4
Administration of estrogens
A
volume of 150 L of an estrogen preparation (0.375 mg Estradiol. benz.,
1.5 mg Estradiol.
phenylpropion; Dimenformon ProlongatumR, N. V. Organon, Oss,
Holland) was injected ip of over 11 weeks old male mice. The mice were
followed for seven days, after which they were sacrificed with CO2.
The testes were collected and frozen in liquid nitrogen.
2.5
Vasectomy
Mice
(n=12) were vasectomized through a mid-abdominal incision under
pentobarbital anaesthesia. The vasa were exposed and ligated using 5-0
Dermalon sutures at two
sites 5 mm from each other and cut between the sutures with scissors.
Sham-operated animals were handled similarly except that the vasa were
left intact. The abdominal wound was closed using 5-0 Dermalon sutures.
The mice were followed for
17, 35, 52 and 70 days, after which they were sacrificed with CO2.
The testes were collected and frozen in liquid nitrogen.
2.6
Cryptorchidism
Mice
(n=3) were made unilaterally cryptorchid through mid-abdominal
incision under pentobarbital (MebunatR, Orion, Turku, Finland) anaesthesia.
The left testis was drawn from the scrotum to the abdominal cavity and
sutured to the parietal peritoneum with 5-0 Dermalon sutures. Sham*-operated
animals were handled similarly except that both testes were left intact.
The mice were followed for seven days, after which they were sacrificed
with CO2. The testes were collected and frozen in liquid nitrogen
for later use.
2.7
Varicocele
The
left spermatic vein of three mice was ligated via a mid-abdominal incision
under pentobarbital anaesthesia. Sham-operated mice were handled similarly
except that the spermatic
vein was left intact. The abdominal wound was closed in one layer using
5-0 Dermalon sutures. The mice were followed for 14 days, after which
they were sacrificed with CO2. The testes were collected and
frozen in liquid nitrogen for later use.
2.8
Indirect immunofluorescence (IF)
Frozen
sections 2-5 m in thickness were cut in a cryostat. They were air-dried
briefly at room temperature and then fixed in cold acetone (-20)
for 1-2 minutes. The sections were stored at -20
and just before use, they were soaked
in PBS. The non-specific binding sites were blocked by incubating the
section in 5% normal rabbit serum in PBS (pH 7.4) for 15-20 minutes. After
washing in PBS, blocking of non-specific binding sites was continued with
5% normal rat serum in PBS for 15-20 minutes. The sections were washed
in PBS. Then they were incubated with the primary antibodies (diluted
into the concentration of 2.0 g/mL in 1% BSA in PBS; Sigma, St. Louis,
MO) for 60 minutes. The sections were washed in PBS and incubated for
60 minutes with the secondary antibody (FITC-conjugated rabbit-anti-rat
Ig, Dako, cat no. F 0234, Copenhagen, Denmark, diluted in 5% normal mouse
serum in 1% BSA in PBS at the concentration of 10 g total protean/mL).
After washing in PBS the sections were mounted in 1,4-diazabicyclo2.2.2.octane
(DABCO, Sigma, St. Louis, MO) -containing glycerol (50% glycerol in 2PBS,
0.1% NaN3 and 100 mg/mL DABCO). The sections were examined
and photographed under an ultraviolet-microscope
equipped with an epi-illuminator and appropriate filters (Leitz, Wetzlar,
Germany).
2.9
Double immunocytochemistry
3 Results
3.1
IL-6
IL-6
was detected in the testes of postpubertal intact mice, mice treated for
70 days
with testosterone or empty implants and cryptorchid and sham-operated
mice. IL-6 is localised to the Leydig cells (Figs.
1a and b). Of the 77 mice studied, 15 (19.5%) had IL-6 expressing
cells at least in the other testis. There were no differences in the expression
of IL-6 between the testosterone and empty implant-treated mice or between
the cryptorchid and sham-operated mice.
3.2
IL-10
IL-10+
cells were present in all the studied testes except in two mice treated
with testosterone for 17 days, in all three mice vasectomized for 52 days,
in one mouse
each in the 14, 35 and 52 days sham-operated groups as well as in all
the three mice in
the varicocele group. Thus, 66 mice out of 77 (85.7%) investigated were
positive for IL-10. IL-10 was localised at the interstitial cells (Figures.
1c-h). Two types of IL-10+ cells were present, 1) morphologically
macrophage (m)-like cells, 2) other
interstitial cells, probably Leydig cells. The latter often showed weaker
staining intensity than the m-like
IL-10+ cells. The identity of the IL-10+ morphologically m-like cells
was investigated using double immunocytochemistry. The IL-10+ m-like
cells were found in several treatment groups: prepubertal intact mice,
postpubertal intact mice (11 weeks old), mice treated for 52 days with
testosterone or empty implant, mice treated for 35, 52 and 70 days with
an empty implant, mice treated 70 days with a testosterone implant, 35
and 70 days vasectomized and sham operated (vasectomy control) mice, mice
treated for 7 days
with estrogen, and 14 days sham operated (varicocele control) mice. In
30 mice out of 77 (39.0%) IL-10+ m cells were detected in the interstitial
tissue of the testis (Table 1). Double staining was made to identify the
IL-10+, morphologically m-like cells.
IL-10 colocalized with the macrophage-specific CD11b in the testes of
intact 9-11 weeks old mice (Figure 2).
As the normal mouse testis does not contain granulocytes, the IL-10+ interstitial
cells were thus most probably macrophages.
3.3
Other cytokines and controls
Figure
1. a) IL-6 in interstitial tissue of a mouse treated
with a testosterone implant for 35 days; immunocytochemistry on a frozen
section (1267), b) the samefield as in Figure 1a, a phase contrast (1267),
c) IL-10+ cells (arrows) in interstitial tissue of a mouse treated with
an empty implant for 52 days; immunocytochemistry on a frozen section
(1267), d) the same field as in Figure 1c, phase contrast
(1267), e) IL-10+ cells (arrows) in interstitial tissue of amouse treated
with an empty implant for 52 days; immunocytochemistry on a frozen section
(633), f) the same field as in Figure 1e, phase contrast (633), g)
IL-10 in the interstitial
tissue of a mouse treated with a testosterone implant for 70 days; immunocytochemistry
on a frozen section (1267), h) the same field as in Figure 1g,
phase contrast (1267), i) CD106 in the interstitial tissue of a mouse
testis 52 days after vasectomy; immunocytochemistry on a frozen section
(1267),j) the same field as in figure 1i, phase contrast (1267), k)
IL-4 staining of a
testis of a mouse treated with an empty implant for 52 days, IL-4 is not
present; immunocytochemistry
on frozen section (1267), l) the same field as in Figure1k, phase contrast
(1267).
Figure 2. IL-10 immunocytochemistry
on frozen section (a and d) as well as CD11b (b and
e), the same field in c as in a and b, the same field in f as in d and
e. Figure 1a-c 1267,
Figure 1d-f 633.
Figure
3. A simplified illustration of immunological microenvironment of
testis. If a T cell
migrating into testicular interstitial tissue meets its antigen there
on MHC-II of testicular macrophages (M), it should go anergic because
of lack of necessary co-stimulating factors CD80 and CD86[60]. In addition
to lack of necessary co-stimulation, clonal anergy is promoted by secretion
of IL-10 by test icular
interstitial cells as shown in present study. Furthermore, TGF-1 produced by
Sertoli cells affects Tcell activation in this site. FasL affects only
activated T cells.
Table 1. Presence of IL-10+ Leydig cells, IL-10+ macrophage-like cells, IL-6, IL-2, IL-4, IL-12, TNF- and IFN- in testes of mice after various treatments.
|
Treatment
groups |
IL-10+ |
IL-10+
m |
IL-6 |
IL-2 |
IL-4 |
IL-12 |
TNF- |
IFN- |
|
Intact
mice: |
||||||||
|
Prepubertal.
(10 d) |
4/4 |
2/4 |
0/4 |
0/4 |
0/4 |
0/4 |
0/4 |
0/4 |
|
Pubertal.
(4 w) |
3/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Postpub.
(9-11 w) |
3/3 |
3/3ad |
3/3adg |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Postpub.
(26 w) |
4/4 |
0/4 |
0/4 |
0/4 |
0/4 |
0/4 |
0/4 |
0/4 |
|
Testosterone
implant: |
||||||||
|
17
d |
1/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
35
d |
3/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
52
d |
3/3 |
0/3v |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
70
d |
3/3 |
3/3jmp |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Empty
implant: |
||||||||
|
17
d |
3/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
35
d |
3/3 |
1/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
52
d |
3/3 |
3/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
70
d |
3/3 |
3/3j |
3/3jmp |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Vasectomized: |
||||||||
|
17
d |
3/3p |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
35
d |
3/3p |
2/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
52
d |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
70
d |
3/3p |
3/3jp |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Sham
operated: |
||||||||
|
17
d |
3/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
35
d |
2/3 |
1/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
52
d |
2/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
70
d |
3/3 |
3/3jp |
0/3jp |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Varicocele
(14 d) |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Sham
operated (14 d) |
2/3 |
2/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Cryptorchidism(7
d) |
3/3 |
0/3 |
3/3s |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Sham
operated (7 d) |
2/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
|
Estrogen
treated(7 d) |
3/3 |
2/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
0/3 |
4 Discussion
The
present results suggest that IL-10 is constitutively expressed in the
interstitial tissue of the testis. As IL10 induces clonal anergy[24,25]
and inhibits TH1, TH2[26] and TH0[27]
mediated immune responses, the present results on constitutive expression
of IL-10 in the testis indicate that IL-10 may have an essential role
in the formation and maintenance of the immune privilege of the testis. Interestingly,
such a role for IL-10 and TGF in the maintenance of the immune privilege
in the anterior chamber of the eye has recently been suggested[28].
On the other hand,
it has been reported that in active autoimmune thyroiditis the IL-10 levels
are significantly higher than in non-autoimmune glands, but in such conditions
the role of IL-10 might be directed to the stimulation of B cell proliferation
and antibody production rather than to the suppression of proinflammatory
cytokine release[29]. Further studies will show the role of
IL-10 in various autoimmune conditions.
It
has been[30] recently confirmed the existence of an immunosuppressive
activity in the interstitial fluid (IF) of the normal rat testis[31].
Their results on the immunosuppressive activity in the IF of the normal
rat testis suggested that the activity could not be neutralised using
a polyspecific TFG antibody. In
addition, it has been found earlier[8,32] that the normal lymphocyte and
leukaemic lymphoblast proliferation inhibiting activity in tissue extracts
of whole abdominal
testes can be neutralised with antibodies neutralising TGF-1. As found
in 1991[33], in the whole abdominal testis extracts, the lymphocyte
3H-TdR incorporation inhibiting activity is higher than in
the scrotal testis and in these conditions
of the abdominal testis a 25 K peak of lymphocyte 3H-TdR incorporation
inhibiting activity appears in the testis in gel filtration chromatography.
It seems that different factors may be involved in immunoregulation in
the normal and abdominal
testis. Furthermore, the responses of immune cells depend also on the
relative roles of TGF and Fas ligand in T-cell regulation[34].
This is especially noteworthy as TGF (25 K) and the soluble form of
Fas ligand (24 K) are localised at the same gel filtration fraction in
the experimental setting[33]. In the present study, the expression
of IL-10 did not change in the abdominal testis, suggesting that IL-10
may not contribute to the previously observed higher
In
contrast to the unchanged IL-10 expression in the abdominal testis, IL-6
was
It
could have been expected on the basis of the recent observations on the
formation of testicular mononuclear cell infiltrates after vasectomy[38]
that in the testes of vasectomized mice the expression of TH1
cytokines would have increased. However, this had not occurred. In further
analysis of the literature, it is evident that vasectomy in fact may promote
humoral immune responses against sperm
autoantigens since only 18% of patients with agenesis or obstruction of
the vas deferens[39] and 32% of vasectomized patients[40]
have a positive leukocyte migration test, but 20-86% of vasectomized patiens
have antisperm antibodies
The
identity of the intensely stained morphologically macrophage-like I
It
has been previously found[51] that IL-10 inhibits the antigen-presenting
capacity of synovial fluid macrophages from patients with rheumatoid arthritis.
In the testis IL-10 secreted by macrophages could significantly down regulate
One
interesting feature of the present results is the observation that i
The
disappearance of IL-10 from the testes of mice with experimental varicocele
is interesting, because this may decrease the local immunosuppression
in the testes and may reflect other changes in testicular macrophage and
Leydig cell
IL-6+
cells were found only in 19.5% of all the testes investigated. Those positive
reactions were not as strong as those for IL-10. IL-6 has been reported
to be produced by human Leydig and Sertoli cells[5] as well
as by mouse Sertoli cells in vitro[54]. In the present
in vivo study, IL-6 was found only in the interstitial tissue. The discrepancy
between our finding, i.e., Sertoli cells of normal testes were IL-6 negative,
and some earlier reports[6,7,54-56] could be explaned that
the majority of those studies have been done in vitro and there
As
IL-6 is capable of inhibiting macrophage pro-inflammatory cytokine production[57],
it is not excluded that IL-6 serves as a complementary cytokine along
with IL-10 in the down-regulation of the lymphocytes and in the creation
of the immunologically privileged site in the testis. Although the present
results do not
allow to make any such conclusions, they encourage to investigate the
role of IL-6 in the testis more thoroughly in the future.
It
has recently been reported that IL-12 is produced by cultured Leydig cells[58].
As IL-12 was never found in the mouse testis in vivo in the present
study, it seems that the Leydig cells start to express IL-12 only in the
extreme unphysiological conditions.
Expression
of IFN- has been reported[13]. In the present study, IFN-
was never found in
the mouse testis, suggesting that its expression by the testicular cells
is a sign of extreme conditions or alternatively, there may be species
differences between the mouse and the rat.
TNF-
production by isolated spermatogenic cells has been reported[14],
but in the present study no TNF- expression in the testis was found
using immunohistochemistry. There may be two reasons for this: 1) differences
between in vivo and
in vitro conditions, 2) differences in the methods used in detection.
Most probably, the in vitro condition of the previous study might
induce abnormal gene
expression.
Acknowledgements
References
[1]
Pöllänen P, Cooper TG. Immunology of the testicular excurrent
ducts. J Reprod Immunol 1994; 26: 167-216.
[2] Hales DB, Diamer T, Hales H. Role of cytokines in testicular function.
Endocrine 1999; 3: 201-17.
[3] Khan SA, Schmidt K, Hallin P, Di Pauli R, De Geyter C, Nieschlag E.
Human testis cytosol and ovarian fluid contain high amounts of interleukin-1-like
factor(s). Mol Cell Endocrinol 1988; 58: 221-30.
[4] Syed V, Söder O, Arver S, Lindh M, Khan S, Ritzen EM. Ontogeny
and cellular o
[5] Cudicini C, Lejeune H, Gomez E, Bosmans E, Ballet F, Saez J, Jgou
B. Human Leydig cells and Sertoli cells are producers of interleukins-1
and -6. J Clin Endocrinol Metab 1997; 82: 1426-33.
[6] Stephan JP, Syed V, Jgou B. Regulation of sertoli cell IL-1 and
IL-6 production in vitro. Moll Cell Endocrinol 1997; 134: 109-18.
[7] Sundström J, Veräjänkorva E, Salminen E, Pelliniemi
LJ, Pöllänen P. Experimental testicular teratoma promotes formation
of humoral immune responses in the host testis. J Reprod Immunol 1999;
42: 107-26.
[8] Pöllänen P, von Euler M, Jahnukainen K, Saari T, Parvinen
M, Sainio-Pöllänen
[9] Teerds KJ, Dorrington JH. Localization of transforming growth factor
beta 1 and beta 2 during testicular development in the rat. Biol Reprod
1993; 48: 40-5.
[10] Tompkins AB, Hutchinson P, deKratser DM, Hedger MP. Characterization
of lymphocytes in the adult rat testis by flow cytometry: Effects of activin
and transforming growth factor beta on lymphocyte subsets in vitro. Biol
Reprod 1998; 58: 943-51.
[11] Lee J, Richburg JH, Younkin SC, Boekelheide K. The Fas system is
a key regulator of germ cell apoptosis in the testis. Endocrinology 1997;
138: 2081-8.
[12] Du X, Everett ET, Wang G, Lee WH, Yang Z, Williams DA. Murine interleukin-11
(IL-11) is expressed at high levels in the hippocampus and expression
is developmentally regulated in the testis. J Cell Physiol 1996; 168:
362-72.
[13] Dejucq N, Dugast I, Ruffault A, van der Meide PH, Jgou B. Interferon-alpha
and -gamma expression in the rat testis. Endocrinology, 1995; 136: 4925-31.
[14] De SK, Chen HL, Pace JL, Hunt JS, Terranova PF, Enders GC. Expression
of tumor necrosis factor-alpha in mouse spermatogenic cells. Endocrinology
1993; 133, 389-96.
[15] Jones R, Dott HM. Changes in luminal plasma and disappearance of
spermatozoa from the
ligated cauda epididymis of the androgen-deficient rabbit. J Reprod Fertil
1980; 60: 65-72.
[16] Pelletier RM. Blood barriers of the epididymis and vas deferens act
asynchronously with the blood barrier of the testis in the mink. Microsc
Res Techn 1994; 27:
333-49.
[17] Huang HF, Nieschlag E. Suppression of the intratesticular testosterone
is associated with quantitative changes in spermatogonial populations
in intact adult rats.
Endocrinology 1986; 118: 619-27.
[18] Tung KSK, Ellis LE, Childs GV, Dufau M. The dark mink: a model of
male infertility. Endocrinology 1984; 114: 922-9.
[19] Kerr JB, Risbridger GP, Murray PJ, Knell CM. Effect of unilateral
cryptorchidism on the intertubular tissue of the adult rat testis: evidence
for intracellular changes within the Leydig cells. Int J Androl 1988;
11: 209-23.
[20] Jarow JP, Budin RE, Dym M, Zirkin BR, Noren S, Marshall FF. Quantitative
pathologic changes in the human testis after vasectomy. A controlled study.
N EnglJ Med 1985; 313:
1252-6.
[21] Flickinger CJ, Herr JC, Howards SS, Caloras D, Yarbo ES, Spell DR,
Gallien TN. The influence of vasovasostomy on testicular alternations
after vasectomy in Lewis rats. Anat Rec 1987; 217: 137-45.
[22] Ylinen L, Teros T, Liukas A, Arvilommi P, Sainio-Pöllänen
S, Veräjänkorva E, Pöllänen
P, Simell O. The role of lipid antigen presentation, cytokine balance
and major histocompatibility in a novel murine model of adoptive transfer
of insulitis. Pancreas 2000; 20: 197-205.
[23] Sainio-Pöllänen S, Liukas A, Simell O, Pöllänen
P. The role of CD8+ cells, cell degeneration and Fas ligand in insulinitis
after intraperitoneal transfer of NOD splenocytes.
Pancreas 1998; 18: 282-93.
[24] Enk AH, Angeloni VL, Udey MC, Katz SI. Inhibition of Langerhans cell
antigen-presenting function by IL-10. A role for IL-10 in induction of
tolerance. J Immunol 1993; 151: 2390-8.
[25] Becker JC, Czerny C, Brocker, EB. Maintenance of clonal anergy by
endogenously produced IL-10. Int Immunol 1994; 6: 1605-12.
[26] Del Prete G, De Garli M, Almerigogna F, Giudizi MG, Biagiotti R,
Romagnani S. Human IL-10 is produced by both type 1 helper (Th1) and type
2 (Th2) cell clones and inhibits their antigen-spesific proliferation
and cytokine production. J Immunol
1993; 150: 353-60.
[27] Silva-Teixeira DN, Contigli C, Goes AM. Cytokine profile associated
to effector functions of human T cell clones specific for Schistosoma
mansoni antigens. Hum Immunol 1998; 59: 219-24.
[28] D'Orazio TJ, Niederkorn JY. A novel role for TGF beta and IL-10
in the induction of immune privilege. J Immunol 1998, 160: 2089-98.
[29] De la Vega JR, Vilaplana JC, Biro A, Hammond L, Bottazzo GF, Mirakian
R. IL-10 expression in thyroid glands: protective or harmful role against
thyroid autoimmunity? Clin Exp Immunol 1998; 113: 126-35.
[30] Hedger MP, Niolic-Paterson DJ, Hutchinson P, Atkinson RC, de Kretser
DM. Immunoregulatory activity in adult rat testicular interstitial fluid:
Roles of interleukin-1 and transforming growth factor beta. Biol Reprod
1998; 58: 927-34.
[31] Pöllänen P, Söder O, Uksila J. Testicular immunosuppressive
protein. J Reprod Immunol 1988; 14: 125-38.
[32] Jahnukainen K, Atterno P, Saari T, Salmi TT, von Euler M, Pöllänen
P, Söder O. Effects of testicular cytokines on proliferation of rat
T-leukaemic lymphoblasts in vitro. J Reprod Immunol 1995; 28: 263-73.
[33] Sainio-Pöllänen, S, Pöllänen P, Setchell BP.
Testicular immunosuppressive activity
in experimental hypogonadism and cryptorchidism. J Reprod Immunol 1991;
20: 59-72.
[34] Chen J\|J, Sun Y, Nabel GJ. Regulation of the proinflammatory effects
of fas ligand (CD95L). Science 1998; 282: 1714-7.
[35] Rost A, Ehrenberg W, Fiedler U. Determination of auto-antibodies
against spermatozoa in experimental cryptorchism. Urol Int 1981; 36: 59-66.
[36] Draehmpaehl D. The behavior of sperm antibody titers after immunization
of guinea pigs with their own sperm and after experimental unilateral
cryptorchism. Berl Mnch Tierarztl Wochenschr 1994; 107: 157-62.
[37] Urry RL, Carrell DT, Starr NT, Snow BW, Middleton RG. The incidence
of antisperm antibodies in infertility patients with a history of cryptorchidism.
J Urol 1994; 151:
381-3.
[38] Aitken H, Kumarukuru S, Orr R, Reid O, Bennett NK, McDonald SW. Effect
of long-term vasectomy on seminiferous tubules in the guinea pig. Clin
Anat 1999; 12: 250-63.
[39] Dondero F, Lenzi A, Picardo M, Pastore R, Valesini G. Cell-mediated
antisperm immunity in selected forms of male infertility. Andrologia 1980;
12: 25-9.
[40] Nagarkatti PS, Rao SS. Cell-mediated immunity to homologous spermatozoa
following vasectomy in the human male. Clin Exp Immunol 1976; 26: 239-42.
[41] Phadke AH, Padukone. Presence and significance of autoantibodies
against spermatozoa in the blood of men with obstructed vas deferens.
J Reprod Fertil 1964; 7:
163-70.
[42] Ansbacher R, Keung-Yeung K, Wurster JC. Sperm antibodies in vasectomized
men. Fertil Steril 1972; 23: 640-3.
[43] Shulman S, Zappi E, Ahmed U, Davis JE. Immunologic consequences of vasectomy.
Contraception 1972; 5: 269-78.
[44] Linnet L, Hjort T. Sperm agglutinins in seminal plasma and serum
after vasectomy: correlation between immunological and clinical findings.
Clin Exp Immunol 1977; 30: 413-20.
[45] Linnet L, Fogh-Andersen P. Vasovasostomy: sperm agglutinins in operatively
obtained epididymal fluid and in seminal plasma before and after operation.
J Clin Lab Immunol 1979; 2: 245-8.
[46] Fuchs EF, Alexander NJ. Immunologic considerations before and after
vasovasostomy. Fertil Steril 1983; 40: 497-9.
[47] Bronson R, Cooper G, Rosenfeld D. Sperm antibodies: their role in
infertility. Fertil Steril 1984; 42: 171-83.
[48] Meinertz H, Linnet L, Fogh-Andersen P, Hjort T. Antisperm antibodies
and fertility after vasovasostomy: a follow-up study of 216 men. Fertil
Steril 1990; 54: 315-21.
[49] Pöllänen P, Maddocks S. Macrophages, Lymphocytes and MHC
II antigen in the ram and the rat testis. J Reprod Immunol 1988; 82: 437-45.
[50] Kern S, Robertson SA, Mau VJ, Maddocks S. Cytokine secretion by macrophages
in the rat testis. Biol Reprod 1995: 53: 1407-16.
[51] Möttönen M, Isomäki P, Saario R, Toivanen P, Punnonen
J, Lassila O. Interleukin-10 inhibits the capasity of synovial macrophages
to function as antigen-presenting cells. Br J Rheumatol 1998; 37: 1207-14.
[52] Yule TD, Montoya GD, Russell LD, Williams TM, Tung KS. Autoantigenic
germ cells exist outside the blood testis barrier. J Immunol 1988; 141:
1161-7.
[53] Zalata A, Hafez T, van Hoecke MJ, Comhaire F. Evalution of beta-endorphin
and interleukin-6 in seminal plasma of patients with certain andrological
diseases. Hum Reprod 1995; 10: 3161-5.
[54] Riccioli A, Filippini A, De Ceasaris P, Barbacci E, Stefanini M,
Starace G, Ziparo E. Inflammatory mediators increase surface expression
of integrin ligands, adhesion to lymphocytes, and secretion of interleukin
6 in mouse Sertoli cells. Proc Natl Acad Sci 1995; 92: 5808-12.
[55] De Cesaris P, Starace D, Riccioli A, Padula F, Filippini A, Ziparo
E. Tumor necrosis factor--alpha induces interleukin-6 production and integrin
ligand expression by distinct transduction pathways. J Biol Chem 1998;
273: 7566-71.
[56] Guitton N, Brouazin-Jousseaume V, Dupaix A, Jegou B, Chenal C. Radiation
effect on rat Sertoli cell function in vitro and in vivo. Int
J Radiat Biol 1999;
75: 327-33.
[57] Ulich TR, Yin S, Guo K, Yi ES, Remick D, del Castillo J. Intratracheal
injection of endotoxin and cytokines. II. Interleukin-6 and transforming
growth factor beta inhibit acute inflammation. Am J Pathol 1991; 138:
1097-101.
[58] Valenti S, Villaggio B, Cutolo M, Giusti M, Giardano G. Preluminary
data suggesting production of interleukin-12 by rat Leydig cells cultured
in vitro. Ann
NY Acad Sci 1999; 876: 259-61.
[59] Li H, Ren J, Dhabuwala CB, Schichi H. Immunotolerance induced by
intratesticular antigen
priming: Expression of TGF, fas and fas ligand. Ocular Immunol Infl
1997; 5: 75-84.
[60] Sainio-Pöllänen
S, Saari T, Simell O, Pöllänen
P. CD28-CD86 interactions
in testicular immunoreculation. J Reprod Immunol 1996; 31: 145-63.
Correspondence
to: BcM
Esko Veräjänkorva, University of Turku, Institute of Biomedicine,
Department of Anatomy, Kiinamyllynkatu 10, FIN-20520 Turku, Finland.
Tel: +358-2-333 7364 Fax: +358-2-333 7352
e-mail: esolve@utu.fi
Received
2000-12-18 Accepted 2001-02-22
